Symbiotic skin bacteria as a source for sex-specific scents in frogs

Amphibians are known to possess a wide variety of compounds stored in their skin glands. While significant progress has been made in understanding the chemical diversity and biological relevance of alkaloids, amines, steroids, and peptides, most aspects of the odorous secretions are completely unknown. In this study, we examined sexual variations in the volatile profile from the skin of the tree frog Boana prasina and combined culture and culture-independent methods to investigate if microorganisms might be a source of these compounds. We found that sesquiterpenes, thioethers, and methoxypyrazines are major contributors to the observed sex differences. We also observed that each sex has a distinct profile of methoxypyrazines, and that the chemical origin of these compounds can be traced to a Pseudomonas sp. strain isolated from the frog´s skin. This symbiotic bacterium was present in almost all individuals examined from different sites and was maintained in captive conditions, supporting its significance as the source of methoxypyrazines in these frogs. Our results highlight the potential relevance of bacteria as a source of chemical signals in amphibians and contribute to increasing our understanding of the role that symbiotic associations have in animals.Fil: Brunetti, Andrés Eduardo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Biología Subtropical. Instituto de Biología Subtropical - Nodo Posadas | Universidad Nacional de Misiones. Instituto de Biología Subtropical. Instituto de Biología Subtropical - Nodo Posadas; Argentina. Universidade de Sao Paulo; BrasilFil: Lucio Lyra, Mariana. Universidade Estadual Paulista Julio de Mesquita Filho; BrasilFil: Melo, Weilan G. P.. Universidade de Sao Paulo; BrasilFil: Andrade, Laura Elena. Universidade de Sao Paulo; BrasilFil: Palacios Rodríguez, Pablo. Universidad de los Andes; ColombiaFil: Prado, Bárbara M.. Universidade de Sao Paulo; BrasilFil: Baptista Haddad, Célio Fernando. Universidade Estadual Paulista Julio de Mesquita Filho; BrasilFil: Tallarico Pupo, Monica. Universidade de Sao Paulo; BrasilFil: Peporine Lopes, Norberto. Universidade de Sao Paulo; Brasi

Multiple origins of green coloration in frogs mediated by a novel biliverdin-binding serpin

Many vertebrates have distinctive blue-green bones and other tissues due to unusually high biliverdin concentrations—a phenomenon called chlorosis. Despite its prevalence, the biochemical basis, biology, and evolution of chlorosis are poorly understood. In this study, we show that the occurrence of high biliverdin in anurans (frogs and toads) has evolved multiple times during their evolutionary history, and relies on the same mechanism—the presence of a class of serpin family proteins that bind biliverdin. Using a diverse combination of techniques, we purified these serpins from several species of nonmodel treefrogs and developed a pipeline that allowed us to assemble their complete amino acid and nucleotide sequences. The described proteins, hereafter named biliverdinbinding serpins (BBS), have absorption spectra that mimic those of phytochromes and bacteriophytochromes. Our models showed that physiological concentration of BBSs fine-tune the color of the animals, providing the physiological basis for crypsis in green foliage even under near-infrared light. Additionally, we found that these BBSs are most similar to human glycoprotein alpha-1-antitrypsin, but with a remarkable functional diversification. Our results present molecular and functional evidence of recurrent evolution of chlorosis, describe a biliverdin-binding protein in vertebrates, and introduce a function for a member of the serpin superfamily, the largest and most ubiquitous group of protease inhibitors.Fil: Taboada, Carlos Alberto. University of Duke; Estados Unidos. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Museo Argentino de Ciencias Naturales "Bernardino Rivadavia"; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química, Física de los Materiales, Medioambiente y Energía. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química, Física de los Materiales, Medioambiente y Energía; Argentina. Universidade de Sao Paulo; BrasilFil: Brunetti, Andrés Eduardo. Universidade de Sao Paulo; Brasil. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Biología Subtropical. Instituto de Biología Subtropical - Nodo Posadas | Universidad Nacional de Misiones. Instituto de Biología Subtropical. Instituto de Biología Subtropical - Nodo Posadas; ArgentinaFil: Lucio Lyra, Mariana. Universidade Estadual Paulista Julio de Mesquita Filho; Brasil. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Fitak, Robert R.. University of Duke; Estados Unidos. University of Central Florida; Estados UnidosFil: Faigon Soverna, Ana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Museo Argentino de Ciencias Naturales "Bernardino Rivadavia"; ArgentinaFil: Ron, Santiago R.. Pontificia Universidad Católica del Ecuador; EcuadorFil: Lagorio, María Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química, Física de los Materiales, Medioambiente y Energía. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química, Física de los Materiales, Medioambiente y Energía; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Inorgánica, Analítica y Química Física; ArgentinaFil: Baptista Haddad, Célio Fernando. Universidade Estadual Paulista Julio de Mesquita Filho; BrasilFil: Peporine Lopes, Norberto. Universidade de Sao Paulo; BrasilFil: Johnsen, Sönke. University of Duke; Estados UnidosFil: Faivovich, Julián. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Museo Argentino de Ciencias Naturales "Bernardino Rivadavia"; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; ArgentinaFil: Chemes, Lucia Beatriz. Universidad Nacional de San Martín; ArgentinaFil: Bari, Sara Elizabeth. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química, Física de los Materiales, Medioambiente y Energía. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química, Física de los Materiales, Medioambiente y Energía; Argentin

Mitochondrial and morphological variation populations of New World Screwworm fly, Cocchliomyia hominivorax

Orientadores: Ana Maria Lima de Azeredo-Espin, Louis Bernard LarczkoTese (doutorado) - Universidade Estadual de Campinas, Instituto de BiologiaResumo: A mosca da bicheira, Cochliomyia hominivorax, é um ectoparasita de animais de sangue quente endêmico da região Neotropical, responsável por causar graves prejuízos à pecuária. Nesta tese, diferentes aspectos da variação molecular e morfológica da espécie foram abordados com o objetivo de se obter um cenário mais amplo sobre a extensão e distribuição da diversidade genética em C. hominivorax. As principais contribuições da tese estão compiladas em duas partes. Uma delas trata da diversidade do DNA mitocondrial (DNAmt) da espécie e está dividida em 4 trabalhos. O primeiro (Artigo 1) trata da diversidade genética e estrutura de populações no Uruguai; os resultados sugerem que as populações nessa localidade são panmíticas e polimórficas com relação ao DNAmt. O segundo trabalho (Artigo 2) apresenta um estudo da distribuição da diversidade mitocondrial em 34 populações de 10 diferentes países, desde o Caribe até o Uruguai. Na região do Caribe foi identificada alta estruturação populacional e baixos índices de diversidade, enquanto que na América do Sul foi identificada alta diversidade e baixa estruturação populacional. Os resultados revelaram um padrão complexo para a distribuição da diversidade genética, sugerindo que diferentes processos estão atuando na formação e manutenção do padrão observado. O terceiro trabalho desta parte trata da caracterização da variação de tamanho da região controle do DNAmt, identificada em alguns indivíduos da espécie; essa variação se deu por repetições em série dos blocos de seqüência conservada e parece ter surgido de forma independente em cada localidade. A análise do domínio B da região controle sugere que este pode ser um potencial marcador para análise de populações, uma vez que revelou grande polimorfismo intra-específico. O quarto trabalho investiga a relação entre as amostras de diferentes regiões do Caribe e América continental por seqüenciamento do gene COII; os resultados sugerem que as populações de Cuba são bem diferenciadas e que as populações do Caribe podem ser originadas de diferentes eventos de colonização. A outra parte das contribuições desta tese é formada por dois trabalhos e trata da caracterização da variação morfológica da asa. O primeiro deles (Artigo 3) caracteriza a variação morfológica de asas de C.hominivorax entre sexos e localidades. Foi identificado um grande dimorfísmo sexual na espécie para tamanho e forma da asa, sugerindo a existência de diferentes pressões seletivas em cada sexo. Também foram identificadas variações entre as populações analisadas, que não estão correlacionadas com as diferenças de latitude e temperatura, sugerindo que diferentes características das localidades, dos hospedeiros ou eventos históricos podem influenciar na variação das asas. O segundo trabalho dessa parte apresenta uma comparação de morfologia da asa entre as espécies C. hominivorax e C. macellaria. Os resultados indicam que análises de morfometria geométrica podem ser utilizadas para identificação das espécies, sendo essa abordagem uma estratégia para a rápida identificação de C. hominivorax e monitoramento de eventos de invasão desta espécie em áreas onde foi erradicada. Os resultados obtidos nos dois trabalhos dessa parte demonstram que a variação da asa é um potencial marcador para estudo de variação morfológica entre populações da mosca da bicheira e entre espécies do gênero CochliomyiaAbstract: The New World screwworm fly, Cochliomyia hominivorax, is an important parasitic insect pest in Neotropical region, and represents a serious threat to the livestock sector. In this thesis we analyzed different aspects of molecular and morphological variation in the species, aiming to obtain a clear scenario about the extent and distribution of genetic diversity in C. hominivorax. The main contributions of this thesis are organized in two parts. The first one, composed of four different works, contains the results about mitochondrial DNA (mtDNA) diversity in the species. In the first work (Article 1), mtDNA was used to study the diversity and population structure of C. hominivorax in Uruguay; results indicated high mtDNA variability and suggested that in Uruguay species form a single panmitic population. In the second work (Article 2) we analyzed 34 populations of C. hominivorax in ten different countries of Caribbean and South America, encompassing almost all the current distribution of the species. Significant population structure and low variability were found in Caribbean populations; in contrast, high variability and low differentiation was found among mainland populations. Our results showed a complex pattern of population genetic structure for the species, suggesting that different processes are acting to the shaping and maintenance of the observed patterns. The third work in this part is about the characterization of length variation in the mtDNA control region; the variation was due to the existence of tandem repetitions of conserved sequence blocks in the control region and the observations suggested that repetitions may be independent events in each locality. The analysis of variable domain revealed high intra-specific polymorphism and suggested that this region may be a potential marker for the study of species populations. The fourth work analyses the relationship between Caribbean and mainland population; Cuban populations differs significantly from all others results suggested different colonization events for Caribbean region. The other part of this thesis consists of two works about characterization of wing variation. The first one (Article 3) provides a morphological analysis of wing variation of C. hominivorax and evaluate the potential use of this morphological character for population variation studies. We found a striking sexual dimorphism in the species for both wing size and shape and suggested that it reflects a history of different selection pressures operating on males and females. We also found morphological variation between populations, which were not associated with latitude or temperature; these preliminary results suggested that C. hominivorax morphology might be affected by locality and/or host characteristics. The second work is about wing variation between C. hominivorax and C. macellaria, results indicated that geometric morphometric methods are useful for species identification and that this strategy could be useful in monitoring invasions events in regions where species have been previous eradicated. Results of this part indicated that wing shape and size may be a reliable marker for population analyses of New World Screwworm, and for inter-specific analyses in genus Cochliomyia.DoutoradoGenetica Animal e EvoluçãoDoutor em Genetica e Biologia Molecula

A lost species or the loss of stripes? The case of Contomastix lizards from Cabo Polonio, Uruguay, with observations on C. lacertoides (Duméril & Bibron) and Cnemidophorus grandensis Cope (Squamata, Teiidae)

The main goal of this manuscript is the reevaluation of the taxonomic status of the teiid lizard Contomastix charrua,known only from Cabo Polonio, a small coastal rocky outcrop in southeastern Uruguay. This species was erected on the basis of the presence of a second pair of ceratobranchials and longer cornua in the hyoid bone, in addition to a reduced expression of the pattern of coloration as compared with C. lacertoides. Nevertheless, we found that both species have indistinguishable hyoid morphology, bearing C. lacertoides a noticeable second pair of ceratobranchials. Besides, we realized that the pattern of coloration in this species is more variable than previously considered. As a result of the present work, C. charrua is included in the synonymy of C. lacertoides. In addition, we provide some observations on the holotype of Cnemidophorus grandensis, a junior synonym of C. lacertoidesFil: Borteiro, Claudio. Museo Nacional de Historia Natural; UruguayFil: Kolenc, Francisco. Museo Nacional de Historia Natural; UruguayFil: Prigioni, Carlos. Museo Nacional de Historia Natural; UruguayFil: Lucio Lyra, Mariana. Universidade Estadual Paulista Julio de Mesquita Filho; BrasilFil: Baldo, Juan Diego. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Nordeste. Instituto de Biología Subtropical. Instituto de Biología Subtropical - Nodo Posadas; Argentin

The Green Clade grows: A phylogenetic analysis of Aplastodiscus (Anura; Hylidae)

Green tree frogs of the genus Aplastodiscus occur in the Atlantic Forest and Cerrado biomes of South America. The genus comprises 15 medium-sized species placed in three species groups diagnosed mainly by cloacal morphology. A phylogenetic analysis was conducted to: (1) test the monophyly of these species groups; (2) explore the phylogenetic relationships among putative species; and (3) investigate species boundaries. The dataset included eight mitochondrial and nuclear gene fragments for up to 6642 bp per specimen. The results strongly support the monophyly of Aplastodiscus and of the A. albofrenatus and A. perviridis groups. Aplastodiscus sibilatus is the sister taxon of all other species of Aplastodiscus, making the A. albosignatus Group non-monophyletic as currently defined. At least six unnamed species are recognized for Aplastodiscus, increasing the diversity of the genus by 40%. A fourth species group, the A. sibilatus Group is recognized. Aplastodiscus musicus is transferred from the A. albofrenatus Group to the A. albosignatus Group, and A. callipygius is considered a junior synonym of A. albosignatus. Characters related to external cloacal morphology reveal an interesting evolutionary pattern of parallelisms and reversions, suggesting an undocumented level of complexity. We analyze, in light of our phylogenetic results, the evolution of reproductive biology and chromosome morphology in Aplastodiscus.Fil: Von Muller Berneck, Bianca. Universidade Estadual Paulista Julio de Mesquita Filho; BrasilFil: Haddad, Célio F.B.. Universidade Estadual Paulista Julio de Mesquita Filho; BrasilFil: Lucio Lyra, Mariana. Universidade Estadual Paulista Julio de Mesquita Filho; BrasilFil: Cruz, Carlos A.G.. Universidade Federal do Rio de Janeiro; BrasilFil: Faivovich, Julián. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Museo Argentino de Ciencias Naturales "Bernardino Rivadavia"; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Biodiversidad y Biología Experimental; Argentin

Ecological niche modeling and new distribution records of the central dwarf frog Physalaemus centralis Bokermann, 1962 (Anura, Leptodactylidae, Leiuperinae) with comments on its taxonomic status

Physalaemus centralis is distributed over open areas of eastern South America. Herein, we perform an extensive review of occurrence records and propose a potential distribution based on ecological niche modeling. In order to confirm species identification, as well taxonomic status along its distribution, we provide morphological and molecular data from analysed specimens. To reach our goals, we compiled data available in the literature and from scientific collections, as well as unpublished data. We identified records of P. centralis in 123 localities, 86 of those records remain unpublished. Consequently, it is possible to extend its range of distribution to the state of Rondonia, Brazil, and also to the Caatinga formation. The Maxent algorithm provided a model of potential distribution consistent with the distribution proposed here. Results indicate that P. centralis has a large distribution associated to open areas of South America (Cerrado, Caatinga and Pantanal), with no evidence that this is a case of species complex. The multiple approach proposed herein, that combines occurrence data from different sources and molecular data to confirm species identification, provided consistent results. Therefore, we recommend that this framework be used as a protocol to assess species distribution in future studies, especially in the case of widely distributed species.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Univ Fed Rio Grande, Inst Ciencias Biol, Lab Vertebrados, Av Italia,Km 8, BR-96203900 Rio Grande, RS, BrazilUniv Estadual Paulista UNESP, Inst Biociencias, Dept Zool, Lab Herpetol, Av 24 A,1515 Bela Vista,Caixa Postal 199, BR-13506970 Rio Claro, SP, BrazilUniv Fed Sao Paulo, Inst Ciencias Ambientais Quim & Farmaceut, Dept Ciencias Biol, Lab Ecol & Sistemat, Rua Prof Artur Riedel 275, BR-09972270 Diadema, SP, BrazilUniversidade Federal de São Paulo (UNIFESP), Inst Ciencias Ambientais Quim & Farmaceut, Dept Ciencias Biol, Lab Ecol & Sistemat, Rua Prof Artur Riedel 275, BR-09972270 Diadema, SP, BrazilCAPES: 338632/2010FAPESP: 2013/50741-7Web of Scienc

Draft genome and multi-tissue transcriptome assemblies of the Neotropical leaf-frog Phyllomedusa bahiana

Amphibians are increasingly threatened worldwide, but the availability of genomic resources that could be crucial for implementing informed conservation practices lags well behind that for other vertebrate groups. Here, we describe draft de novo genome, mitogenome, and transcriptome assemblies for the Neotropical leaf-frog Phyllomedusa bahiana native to the Brazilian Atlantic Forest and Caatinga. We used a combination of PacBio long reads and Illumina sequencing to produce a 4.74-Gbp contig-level genome assembly, which has a contiguity comparable to other recent nonchromosome level assemblies. The assembled mitogenome comprises 16,239 bp and the gene content and arrangement are similar to other Neobratrachia. RNA-sequencing from 8 tissues resulted in a highly complete (86.3%) reference transcriptome. We further use whole-genome resequencing data from P. bahiana and from its sister species Phyllomedusa burmeisteri, to demonstrate how our assembly can be used as a backbone for population genomics studies within the P. burmeisteri species group. Our assemblies thus represent important additions to the catalog of genomic resources available from amphibians.Fil: Andrade, Pedro. Universidad de Porto. Facultad de Ciências. Centro de Investigação em Biodiversidade e Recursos Genéticos; PortugalFil: Lucio Lyra, Mariana. Universidade Estadual Paulista Julio de Mesquita Filho; BrasilFil: Zina, Juliana. Universidade Estadual Do Sudoeste Da Bahia; BrasilFil: Bastos, Deivson F.O.. Universidade Estadual Do Sudoeste Da Bahia; BrasilFil: Brunetti, Andrés Eduardo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Biología Subtropical. Instituto de Biología Subtropical - Nodo Posadas | Universidad Nacional de Misiones. Instituto de Biología Subtropical. Instituto de Biología Subtropical - Nodo Posadas; ArgentinaFil: Pontes Baêta Da Costa, Délio. Universidad de Porto. Facultad de Ciências. Centro de Investigação em Biodiversidade e Recursos Genéticos; Portugal. Universidade Estadual Paulista Julio de Mesquita Filho; BrasilFil: Afonso, Sandra. Universidad de Porto. Facultad de Ciências. Centro de Investigação em Biodiversidade e Recursos Genéticos; PortugalFil: Brunes, Tuliana O.. Universidade de Sao Paulo; BrasilFil: Goulart Taucce, Pedro Paulo. Universidade Estadual Paulista Julio de Mesquita Filho; BrasilFil: Carneiro, Miguel. Universidad de Porto. Facultad de Ciências. Centro de Investigação em Biodiversidade e Recursos Genéticos; PortugalFil: Baptista Haddad, Célio Fernando. Universidade Estadual Paulista Julio de Mesquita Filho; BrasilFil: Sequeira, Fernando. Universidad de Porto. Facultad de Ciências. Centro de Investigação em Biodiversidade e Recursos Genéticos; Portuga

Evaluation of a quality improvement intervention to reduce anastomotic leak following right colectomy (EAGLE): pragmatic, batched stepped-wedge, cluster-randomized trial in 64 countries

Background Anastomotic leak affects 8 per cent of patients after right colectomy with a 10-fold increased risk of postoperative death. The EAGLE study aimed to develop and test whether an international, standardized quality improvement intervention could reduce anastomotic leaks. Methods The internationally intended protocol, iteratively co-developed by a multistage Delphi process, comprised an online educational module introducing risk stratification, an intraoperative checklist, and harmonized surgical techniques. Clusters (hospital teams) were randomized to one of three arms with varied sequences of intervention/data collection by a derived stepped-wedge batch design (at least 18 hospital teams per batch). Patients were blinded to the study allocation. Low- and middle-income country enrolment was encouraged. The primary outcome (assessed by intention to treat) was anastomotic leak rate, and subgroup analyses by module completion (at least 80 per cent of surgeons, high engagement; less than 50 per cent, low engagement) were preplanned. Results A total 355 hospital teams registered, with 332 from 64 countries (39.2 per cent low and middle income) included in the final analysis. The online modules were completed by half of the surgeons (2143 of 4411). The primary analysis included 3039 of the 3268 patients recruited (206 patients had no anastomosis and 23 were lost to follow-up), with anastomotic leaks arising before and after the intervention in 10.1 and 9.6 per cent respectively (adjusted OR 0.87, 95 per cent c.i. 0.59 to 1.30; P = 0.498). The proportion of surgeons completing the educational modules was an influence: the leak rate decreased from 12.2 per cent (61 of 500) before intervention to 5.1 per cent (24 of 473) after intervention in high-engagement centres (adjusted OR 0.36, 0.20 to 0.64; P < 0.001), but this was not observed in low-engagement hospitals (8.3 per cent (59 of 714) and 13.8 per cent (61 of 443) respectively; adjusted OR 2.09, 1.31 to 3.31). Conclusion Completion of globally available digital training by engaged teams can alter anastomotic leak rates. Registration number: NCT04270721 (http://www.clinicaltrials.gov)